植物-菌根真菌联合修复重金属污染土壤

菌根是菌根真菌侵染植物根系后在植物根部形成的共生结构。菌根技术作为一种生物强化技术应用于重金属污染土壤的植物修复已引起研究者的广泛关注。目前大量研究表明菌根能强化植物对重金属的转运、富集及根系稳定化过程,并通过促进营养物质的吸收利用、稳定细胞内氧化还原平衡、调控抗逆性相关基因的表达以及改善根际微生态环境等方式提升寄主植物的抗逆性。本文在介绍菌根真菌在植物修复重金属污染的联合过程中的作用效应及机制的基础上,分析了目前限制该技术应用的瓶颈问题以及未来的研究方向,为植物-菌根真菌联合修复的推广应用提供理论基础。     

植物根际促生菌促生特性研究进展

根际微生物组是决定农作物健康状况的关键因素之一,也是调节农作物与生物和非生物环境相互作用的重要因素。植物根际促生菌(plant growth-promoting rhizobacteria, PGPR)为农作物宿主提供了多种有益作用,通过化学交流以复杂的方式与农作物、土壤相互作用,进而促进农作物生长。本文综述了PGPR对农作物的促生机制、PGPR与农作物的互作及其在农业实践中的应用,并展望了PGPR在农业实践中应用的发展趋势,以期为今后PGPR的应用和研究提供新的思路和理论支撑。     

植物促生菌提高植物重金属耐受性研究进展

近年来植物修复技术因其独特的优势而被广泛关注。许多植物被认为是有价值的利用资源, 然而, 最有实际使用价值的植物对重金属的耐受性有限, 实际应用中变得越来越困难。植物促生菌资源对环境无污染, 具有独特的多样性和巨大的潜力。随着资源的开发和技术的发展, 微生物调控将会使植物修复技术变得更加可行和更有价值。回顾近年来新兴的微生物调控技术, 主要是植物促生菌的筛选、鉴定和应用价值。     

丛枝菌根在植物修复重金属污染土壤中的作用

丛枝菌根（Arbuscular mycorrhizae,AM）是自然界中分布最广的一类菌根,AM真菌能与陆地上绝大多数的高等植物共生,常见于包括重金属污染土壤在内的各种生境中。在重金属污染条件下,AM真菌可以减轻重金属对植物的毒害,影响植物对重金属的吸收和转运,在重金属污染土壤的植物修复中显示出极大的应用潜力。重点介绍了AM真菌对植物重金属耐性的影响及其在植物提取和植物稳定中的应用等方面的进展,讨论了未来研究所面临的任务和挑战。     

石油污染土壤中植物根际促生菌的筛选及特性分析

以1-氨基环丙烷-1-羧酸(ACC)为唯一氮源,从黑龙江省大庆地区石油污染土壤的狼尾草根际土壤中分离筛选出2株产ACC脱氨酶的细菌,F4-1和F4-2。对分离的菌株进行生理生化和16S rDNA序列鉴定,确定F4-1为肠杆菌属(Enterobactersp.),F4-2为克雷伯菌属(Klebsiellasp.)。菌株F4-1的ACC脱氨酶活性为(1.40±0.17)μmolα-KA.(mg Pr.h)-1,高于F4-2的(1.03±0.03)μmolα-KA.(mg Pr.h)-1。随着L-Trp浓度的增加,菌株F4-1和F4-2的吲哚乙酸(IAA)合成量相应增加,总体上看F4-1的IAA合成能力高于F4-2。F4-1合成嗜铁素的能力也高于F4-2。     

产铁载体根际菌在植物修复重金属污染土壤中的应用潜力

产铁载体根际菌(siderophore-producing rhizobacteria,SPR)是一类植物根际促生菌,在植物修复重金属污染土壤中可起重要的辅助作用.本文在综合分析国内外相关研究进展的基础上,阐述了SPR缓解重金属植物胁迫毒害的功能及其提高土壤重金属生物活性的机理,并指出SPR在促进重金属污染土壤中植物的生长、增强植物累积重金属的过程等方面具有较好的应用潜力.对当前研究中存在的SPR提高或降低植物累积重金属的矛盾现象也进行了分析.针对当前研究存在的不足,提出今后应深入研究SPR与植物(尤其是重金属超累积植物)之间的相互作用机理,进一步明确影响铁载体螯合、活化土壤重金属的关键因素,综合考虑铁载体对土壤重金属生物活性和对植物吸收重金属的影响,探索在重金属污染土壤中提高SPR定殖能力的强化方法.     

植物根际促生菌的筛选及鉴定

【目的】植物根际促生菌(PGPR)和植物的互作关系往往不稳定,PGPR菌群有可能提高菌株对野外环境的适应性。为此,本文根据PGPR促生机制的多样性,从不同植物根际土壤进行了PGPR的筛选及鉴定。【方法】首先,按照固氮、解磷、解钾、拮抗6种常见病原真菌,同时能在植物根际定殖为基本初筛标准,然后在实验室条件下测定初筛菌株的多项促生能力(PGP),最后通过生理生化试验和16SrRNA基因序列分析对所筛菌株进行鉴定。【结果】从江苏扬州、盐城等地土壤样品筛选出14株PGPR,具有体外抑菌、产NH3、产IAA、产HCN、产嗜铁素、解磷、溶钾、固氮以及产抗生素等促生能力。分类鉴定结果显示:7株属于假单胞菌属(Pseudomonas)、3株属于类芽孢杆菌属(Paenibacillus)、2株为芽孢杆菌属(Bacillus)、1株为布克霍尔德氏菌属(Burkholderia)、1株为欧文氏菌属(Erwinia)。【结论】所筛细菌具有多种促生能力,且能在根际定殖,为进一步构建多功能PGPR广适菌群提供菌株资源。     

超积累植物与丛枝菌根真菌共生及其联合吸收积累重金属的效应

利用超积累植物进行重金属污染土壤修复是应对全球大面积分布无机污染问题的重要解决方法之一。超积累植物虽然具有超量积累重金属的能力,但其定植、生长和提取功能的发挥都受到重金属胁迫的显著影响。利用丛枝菌根真菌 (Arbuscular mycorrhizal fungi,AMF) 强化超积累植物功能可联合发挥二者的功能优势,提升修复效率、缩短修复周期、保持修复效果的稳定性和长期性,在日益复杂、严峻的重金属污染治理领域具有重要的研究价值和广阔的应用前景。文中首先给出了超积累植物的概念、中国本土首次报道的典型重金属元素超积累植物和能与AMF形成共生体系的超积累植物名录,系统深入地探讨了AMF对超积累植物生长和吸收累积重金属的影响,以及超积累植物+AMF联合吸收积累重金属的效应与作用机制,认为AMF可通过调节根围理化与生物条件、元素平衡状况、生理代谢和基因表达等途径,增强超积累植物吸收积累重金属的效应,超积累植物+AMF构建的共生体系具备联合修复重金属污染生境的潜力。最后指出了超积累植物+AMF共生联合修复技术当前面临的关键问题、发展方向和应用前景。     

丛枝菌根对植物根际逆境的生态学意义

近年来,我国在菌根分子生物学、菌根营养学、菌根分类学和菌根生态学等方面取得了令人瞩目的研究成果,其中对丛枝菌根真菌(AMF)的研究居多。AMF能与大部分陆地植物根系形成共生关系,促进植物生长发育,提高植物抗逆性,在保持生态平衡、保护生态环境等方面发挥重要作用。本文主要从非生物胁迫(干旱胁迫、重金属污染、盐碱胁迫)和生物胁迫(致病菌和线虫侵染)方面介绍了AMF在植物根际逆境中发挥的生态功能及作用机制,提出了该研究领域尚存的不足之处和研究前景,为AMF后续研究提供参考。     

柑橘园土壤中丛枝菌根真菌土著菌株的促生效应研究

柑橘对丛枝菌根(AM)真菌具有较高的依赖性,从柑橘园土壤中分离筛选的高效促生AM真菌菌株具有重要的应用价值。本研究从广东增城柑橘园(酸橘砧‘红江橙’)土壤中分离的4个AM真菌土著菌株对‘红江橙’幼苗的促生效应。结果表明,4个土著菌株分别是Scutellospora属和Glomus属菌株,根系侵染率为12.7%～29.3%;与不接种对照相比,4个土著菌株不同程度地促进‘红江橙’幼苗的株高、生物量和N、P、K养分含量,菌根依赖性达9.4%～37.1%;主成分分析表明,土著菌株ZCSP-D的促生效应达到常用优良菌株Rhizophagus irregularis的水平。     

Contribution of the arbuscular mycorrhizal symbiosis to heavy metal phytoremediation

High concentrations of heavy metals (HM) in the soil have detrimental effects on ecosystems and are a risk to human health as they can enter the food chain via agricultural products or contaminated drinking water. Phytoremediation, a sustainable and inexpensive technology based on the removal of pollutants from the environment by plants, is becoming an increasingly important objective in plant research. However, as phytoremediation is a slow process, improvement of efficiency and thus increased stabilization or removal of HMs from soils is an important goal. Arbuscular mycorrhizal (AM) fungi provide an attractive system to advance plant-based environmental clean-up. During symbiotic interaction the hyphal network functionally extends the root system of their hosts. Thus, plants in symbiosis with AM fungi have the potential to take up HM from an enlarged soil volume. In this review, we summarize current knowledge about the contribution of the AM symbiosis to phytoremediation of heavy metals.     

AMF和PGPR联合修复菲和芘污染土壤的效应

丛枝菌根真菌（arbuscular mycorrhizal fungi,AMF）与根围促生细菌（plant growth-promoting rhizobacteria,PGPR）联合降解有毒有机物、修复污染土壤和促进植物生长的作用倍受关注。本试验旨在探究AMF与PGPR联合降解土壤中菲和芘的效应,以菲和芘1:1混合处理浓度各0、50mg/kg、100mg/kg和150mg/kg下对高羊茅Festuca elata接种AMF根内根孢囊霉Rhizophagus intraradices（Ri）、变形球囊霉Glomus versiforme（Gv）、PGPR荧光假单胞菌Pseudomonas fluorescens Ps2-6、芽孢杆菌Bacillus velezensis Ps3-2、Ri+Ps2-6、Ri+Ps3-2、Gv+Ps2-6、Gv+Ps3-2和不接种对照共36个处理。结果表明,供试AMF增加了PGPR的定殖数量;接种PGPR则显著提高AMF的侵染率。AMF、PGPR或AMF+PGPR处理均显著降低土壤中菲和芘含量,促进植物对土壤中菲和芘的吸收,显著提高高羊茅根系和叶片内的菲和芘含量。在土壤中菲和芘100mg/kg和150mg/kg水平下,Gv与Ps2-6及Ri与Ps2-6能相互促进对土壤中菲和芘的去除效应,其中接种Gv+Ps2-6组合处理的去除率最高,达到95%-98%,土壤中多酚氧化酶、脱氢酶和过氧化氢酶活性显著高于单接种处理和不接种对照,而酸性磷酸酶活性变化则表现为相反趋势。其中以Gv+Ps2-6组合处理的多酚氧化酶活性最高,为0.17mg/g,是不接种对照的1.9倍;脱氢酶和过氧化氢酶活性分别达到1.32µg/(g·h)和1.81mL/g;酸性磷酸酶活性则比不接种对照土壤降低27%-45%;易提取球囊霉素相关土壤蛋白含量和总球囊霉素相关土壤蛋白含量分别是不接种对照的1.6倍和1.5倍。     

Increasing phosphorus removal in willow and poplar vegetation filters using arbuscular mycorrhizal fungi

Fast growing woody species are increasingly used in vegetation filters for wastewater treatment. Their efficiency in phosphorus (P) removal notably depends on plant uptake and storage in aboveground tissues. In this study, Populus NM5 (P. nigra × P. maximowiczii), Salix miyabeana (SX64) and Salix viminalis (5027) were planted in pots to evaluate the influence of colonization by arbuscular mycorrhizal fungi (AMF) Glomus intraradices on P uptake using two different P concentrations in irrigation water. Based on analysis of the foliar and woody components, our results show that the two treatments (inoculation with G. intaradices and P-irrigation) interact differently with total P content. Foliar P content is principally enhanced by the P-irrigation concentration, whereas the mycorrhizal colonization increases stem P content. In the presence of G. intraradices, both S. miyabeana and S. viminalis showed a 33% increase in stem P content. The latter finding is mainly due to an increase in biomass production, without modification of the P concentration, indicating that AMF associations affect P use efficiency. Thus, using arbuscular mycorrhizal fungi for phytoremediation strategies may increase biomass productivity and hence improve pollutant uptake.     

Increased photosynthetic acclimation in alfalfa associated with arbuscular mycorrhizal fungi (AMF) and cultivated in greenhouse under elevated CO2

Medicago sativa L. (alfalfa) can exhibit photosynthetic down-regulation when grown in greenhouse conditions under elevated atmospheric CO₂. This forage legume can establish a double symbiosis with nitrogen fixing bacteria and arbuscular mycorrhizal fungi (AMF), which may increase the carbon sink effect of roots. Our aim was to assess whether the association of alfalfa with AMF can avoid, diminish or delay the photosynthetic acclimation observed in previous studies performed with nodulated plants. The results, however, showed that mycorrhizal (M) alfalfa at the end of their vegetative period had lower carbon (C) discrimination than non-mycorrhizal (NM) controls, indicating photosynthetic acclimation under ECO₂ in plants associated with AMF. Decreased C discrimination was due to the acclimation of conductance, since the amount of Rubisco and the expression of genes codifying both large and small subunits of Rubisco were similar or slightly higher in M than in NM plants. Moreover, M alfalfa accumulated a greater amount of soluble sugars in leaves than NM plants, thus favoring a down-regulation effect on photosynthetic rates. The enhanced contents of sugars in leaves coincided with a reduced percentage of arbuscules in roots, suggesting decreased sink of carbohydrates from shoots to roots in M plants. The shorter life cycle of alfalfa associated with AMF in comparison with the NM controls may also be related to the accelerated photosynthetic acclimation in M plants. Further research is needed to clarify to what extent this behavior could be extrapolated to alfalfa cultivated in the field and subjected to periodic cutting of shoots under climatic change scenarios.     

The influence of Rhizobium and arbuscular mycorrhizal fungi on nitrogen and phosphorus accumulation by Vicia faba

BACKGROUND AND AIMS: The aim of this study was to investigate the effects of the interactions between the microbial symbionts, Rhizobium and arbuscular mycorrhizal fungi (AMF) on N and P accumulation by broad bean (Vicia faba) and how increased N and P content influence biomass production, leaf area and net photosynthetic rate. METHODS: A multi-factorial experiment consisting of four different legume-microbial symbiotic associations and two nitrogen treatments was used to investigate the influence of the different microbial symbiotic associations on P accumulation, total N accumulation, biomass, leaf area and net photosynthesis in broad bean grown under low P conditions. KEY RESULTS: AMF promoted biomass production and photosynthetic rates by increasing the ratio of P to N accumulation. An increase in P was consistently associated with an increase in N accumulation and N productivity, expressed in terms of biomass and leaf area. Photosynthetic N use efficiency, irrespective of the inorganic source of N (e.g. NO3- or N2), was enhanced by increased P supply due to AMF. The presence of Rhizobium resulted in a significant decline in AMF colonization levels irrespective of N supply. Without Rhizobium, AMF colonization levels were higher in low N treatments. Presence or absence of AMF did not have a significant effect on nodule mass but high N with or without AMF led to a significant decline in nodule biomass. Plants with the Rhizobium and AMF symbiotic associations had higher photosynthetic rates per unit leaf area. CONCLUSIONS: The results indicated that the synergistic or additive interactions among the components of the tripartite symbiotic association (Rhizobium-AMF-broad bean) increased plant productivity.     

4株茶树根际促生菌菌株的鉴定及促生作用

【背景】根际促生菌可以促进植物生长、提高植物抗性。茶树根际具有特殊的根土微生物生境,可以获得具促生作用的有益微生物。【目的】探究4株茶树根际促生菌菌株的分类地位及促生作用,筛选优良的根际促生菌菌株。【方法】通过形态、生理生化特征、16S rRNA基因序列同源性比对鉴定4株茶树根际促生菌,采用钼锑抗比色法测定溶磷量,通过比色法测定ACC脱氨酶活性、CAS法测定产铁载体能力、Salkowski法测定产IAA (Indoleacetic acid)的能力进行促生作用研究,通过盆栽实验测试白菜、空心菜、苋菜及水稻的株高及鲜重以分析促生效应。【结果】鉴定KKS-6-N1为放射型土壤杆菌(Agrobacteriumradiobacter), KKS-7-N7为铜绿假单胞菌(Pseudomonas aeruginosa),GD3为Pseudomonashunanensis,GD12为弯曲芽孢杆菌(Bacillusflexus)。固氮菌株KKS-6-N1可产铁载体;固氮菌株KKS-7-N7具有解磷及产铁载体能力,分泌的IAA含量高达101.29mg/L;解钾菌株GD3具溶磷能力,分泌的ACC脱氨酶酶活为8.09μmol/(mg·h),相对铁载体含量为0.31;具固氮解钾性能的菌株GD12分泌的ACC脱氨酶活性为14.46μmol/(mg·h)。盆栽试验表明,4个菌株对白菜、空心菜、苋菜的株高和鲜重均有明显促进作用,尤以GD3效果更甚。【结论】茶树根际促生菌菌株Pseudomonas hunanensis GD3促生作用显著,具有开发成微生物菌肥的潜力。     

Growth promotion and yield enhancement of peanut (Arachis hypogaea L.) by application of plant growth-promoting rhizobacteria

Although plant growth-promoting rhizobacteria (PGPR) have been reported to influence plant growth, yield and nutrient uptake by an array of mechanisms, the specific traits by which PGPR promote plant growth, yield and nutrient uptake were limited to the expression of one or more of the traits expressed at a given environment of plant–microbe interaction. We selected nine different isolates of PGPR from a pool of 233 rhizobacterial isolates obtained from the peanut rhizosphere on the basis of ACC-deaminase activity. The nine isolates were selected, initially, on the basis of germinating seed bioassay in which the root length of the seedling was enhanced significantly over the untreated control. All the nine isolates were identified as Pseudomonas spp. Four of these isolates, viz. PGPR1, PGPR2, PGPR4 and PGPR7 (all fluorescent pseudomonads), were the best in producing siderophore and indole acetic acid (IAA). In addition to IAA and siderophore-producing attributes, Pseudomonas fluorescens PGPR1 also possessed the characters like tri-calcium phosphate solubilization, ammonification and inhibited Aspergillus niger and A. flavus in vitro. P. fluorescens PGPR2 differed from PGPR1 in the sense that it did not show ammonification. In addition to the traits exhibited by PGPR1, PGPR4 showed strong in vitro inhibition to Sclerotium rolfsii. The performances of these selected plant growth-promoting rhizobacterial isolates were repeatedly evaluated for 3 years in pot and field trials. Seed inoculation of these three isolates, viz. PGPR1, PGPR2 and PGPR4, resulted in a significantly higher pod yield than the control, in pots, during rainy and post-rainy seasons. The contents of nitrogen and phosphorus in soil, shoot and kernel were also enhanced significantly in treatments inoculated with these rhizobacterial isolates in pots during both the seasons. In the field trials, however, there was wide variation in the performance of the PGPR isolates in enhancing the growth and yield of peanut in different years. Plant growth-promoting fluorescent pseudomonad isolates, viz. PGPR1, PGPR2 and PGPR4, significantly enhanced pod yield (23–26%, 24–28% and 18–24%, respectively), haulm yield and nodule dry weight over the control in 3 years. Other attributes like root length, pod number, 100-kernel mass, shelling out-turn and nodule number were also enhanced. Seed bacterization with plant growth-promoting P. fluorescens isolates, viz. PGPR1, PGPR2 and PGPR4, suppressed the soil-borne fungal diseases like collar rot of peanut caused by A. niger and PGPR4 also suppressed stem rot caused by S. rolfsii. Studies on the growth patterns of PGPR isolates utilizing the seed leachate as the sole source of C and N indicated that PGPR4 isolate was the best in utilizing the seed leachate of peanut, cultivar JL24. Studies on the rhizosphere competence of the PGPR isolates, evaluated on the basis of spontaneous rifampicin resistance, indicated that PGPR7 was the best rhizoplane colonizer and PGPR1 was the best rhizosphere colonizer. Although the presence of growth-promoting traits in vitro does not guarantee that an isolate will be plant growth promoting in nature, results suggested that besides ACC-deaminase activity of the PGPR isolates, expression of one or more of the traits like suppression of phytopathogens, solubilization of tri-calcium phosphate, production of siderophore and/or nodulation promotion might have contributed to the enhancement of growth, yield and nutrient uptake of peanut.     

Synergistic effects of Arbuscular mycorrhizal fungi and plant growth promoting rhizobacteria in bioremediation of iron contaminated soils

Three Arbuscular mycorrhizal fungi (AMF) from Glomus, Acaulospora and Scutellospora, and four plant growth promoting rhizobacteria (PGPR) isolates related to genera Streptomyces, Azotobacter, Pseudomonas and Paenibacillus were found to be effective in phytoremediation of Fe³⁺ contaminated soil where Pennisetum glaucum and Sorghum bicolor were growing as host plants. Co-inoculation of AMF and PGPR showed better results in comparison to either, AMF and PGPR under pot conditions. Both AMF and PGPR were able to produce siderophores. AMF and PGPR associated to P. glaucum and S. bicolor plants increased the extent of iron absorption. AMF and PGPR combination exhibited superior (p < 0.01) phytoremediation efficiency with P. glaucum compared to S. bicolor. These findings warrant further investigations of these synergistic interactions and large-scale in situ studies for bioremediation of iron-contaminated soils.     

The effect of plant growth-promoting rhizobacteria on the phytoextraction of Cd and Zn by Brassica napus L.

The test strains Bacteroidetes bacterium (Ba), Pseudomonas fluorescens (Pf) and Variovorax sp. (Va) were selected in advance for their in vitro capability for growth promotion of rapeseed in the presence of increased concentrations of Cd, Cu, Pb and Zn in the medium. In the pot experiment, the strains were used for single Ba, Pf, Va or combined Ba + Pf, Ba + Va, Pf + Va, and Ba + Pf + Va inoculation of B. napus growing in contaminated soil from alluvial deposits. The positive effect of bacterial strains on plant growth was observed in vitro, but was not confirmed in situ in the contaminated soil, where the tested strains inhibited biomass production, rather than stimulating it. However, single inoculation with Ba significantly increased the chlorophyll content and K⁺ concentration in the leaves. The inoculation of rapeseed with Ba and Va strains was indicated to be the most promising combination for phytoextraction of Cd and Zn from contaminated soil. Combined inoculation with Pf+Va and Pf + Ba+Va significantly decreased the concentration of heavy metals in the roots of rapeseed. We conclude that suitable combinations of PGPR can control the metal uptake of B. napus, selectively increasing either metal extraction or metal stabilization in the rhizosphere and offering promising applications in soil remediation.     

丛枝菌根真菌对植物繁殖的影响研究进展

丛枝菌根真菌(arbuscular mycorrhizal fungi, AMF)与宿主植物所形成的互惠共生体系是生态系统中广泛分布的共生体系代表类型之一。AMF除能够促进宿主植物生长发育外,也可以对宿主植物的繁殖过程产生多方面影响。研究宿主植物在AMF共生状态下繁殖策略的变化规律,对于深入理解植物繁殖适合度的变化具有重要理论意义。该文综述了AMF对宿主植物繁殖影响的相关研究,包括AMF的侵染对宿主植物繁殖分配、花部特征、虫媒传粉和花期的影响。目前已有研究发现某些AMF能够促进宿主植物增加繁殖资源投入,提高花朵产生的数量或花冠直径,同时增加花粉数量和花蜜量来影响访花昆虫的行为,以及造成开花提前及花期延长,但其作用的具体机制尚不明确,且因宿主植物的差异,并未有完全统一的结论。然而,由于AMF与植物共生的普遍性,其在植物繁殖过程中发挥的重要作用不可忽略。今后除了在以上各方面开展更深入的研究外,还需在AMF对宿主植物繁殖性状的影响机制、AMF共生条件下植物有性繁殖过程中雌雄功能的资源分配,以及对无性繁殖和后代适应性的影响等方面进行更深入的研究。